New species of Hymenoptera associated with galls on Calliandra brevipes Benth. (Fabaceae, Mimosoidea) in Brazil

Four species of Hymenoptera: Tanaostigmodes ringueleti (Brèthes, 1924), T. mecanga sp.nov. (Chalcidoidea, Tanaostigmatidae), Allorhogas taua sp. nov. (Braconidae, Doryctinae) and Eurytoma sp. (Chalcidoidea, Eurytomidae) were reared from two different types of galls of Calliandra brevipes Benth. (Fabaceae, Mimosoidea) in Juiz-de-Fora, Minas Gerais State, Brazil. The two Tanaostigmatidae species are probably the gall inducers; the Braconidae species probably is phytophagous inquiline in round gall type. The two new species are described and illustrated, including their immature stages.

A new species of the fungus-farming ant genus Mycetagroicus Brandão & Mayhé-Nunes (Hymenoptera, Formicidae, Attini)

The fungus-farming ant genus Mycetagroicus Brandão & Mayhé-Nunes was proposed based on three species from the Brazilian "Cerrado": M. cerradensis, M. triangularis and M. urbanus. Here we describe a new species of Attini ant of the genus Mycetagroicus, M. inflatus n. sp., based on two workers collected in eastern Pará State, Brazil. A new key for species identification, comments on differences among species and new geographical distribution data are furnished.

A new species of the genus Pimelerodius Vanin from the Amazon Region, with notes on the geografic distribution of Pimelerodius motacilla (Boheman) (Coleoptera, Curculionidae, Otidocephalini)

Pimelerodius punctiventris sp. nov. (type locality Brazil, Amazonas, Itacoatiara) is described and illustrated. The new taxon is compared with similar species, being distinguished from the other 12 known species of the genus by the presence of punctures in ventrite I. The available published key for identification of species of Pimelerodius is adapted to include the new species. A modification of the generic description of the aedeagus of Pimelerodius is provided, a necessity due to the differences observed in the aedeagus of the new species. The occurrence of P. motacilla (Boheman, 1843) in the Amazon Region, recorded in sympatry with P. punctiventris in Itacoatiara, AM, is discussed and confirmed, based on the study of 41 available specimens.

Database construction and Peptide identification strategies for proteogenomic studies on sequenced genomes.

Since the advent of high-throughput DNA sequencing technologies, the ever-increasing rate at which genomes have been published has generated new challenges notably at the level of genome annotation. Even if gene predictors and annotation softwares are more and more efficient, the ultimate validation is still in the observation of predicted gene product( s). Mass-spectrometry based proteomics provides the necessary high throughput technology to show evidences of protein presence and, from the identified sequences, confirmation or invalidation of predicted annotations. We review here different strategies used to perform a MS-based proteogenomics experiment with a bottom-up approach. We start from the strengths and weaknesses of the different database construction strategies, based on different genomic information (whole genome, ORF, cDNA, EST or RNA-Seq data), which are then used for matching mass spectra to peptides and proteins. We also review the important points to be considered for a correct statistical assessment of the peptide identifications. Finally, we provide references for tools used to map and visualize the peptide identifications back to the original genomic information.

Exposure of a single host (Chrysomya megacephala) (Calliphoridae) to different quantities of female parasitoids (Nasonia vitripennis) (Pteromalidae)

The aim of this study was to verify the duration of the development period, number of parasitoids produced per pupa, parasitism rate and sex ratio of Nasonia vitripennis (Hymenoptera, Pteromalidae), when they were exposed to a single host: Chrysomya megacephala (Diptera, Calliphoridae). One pupa was exposed in glass tubes to different numbers of female parasitoids (1, 3, 5, 7, 9 and 11) during 48 h. Twenty replications/treatment were used, under controlled conditions (T= 27 °C day/ 25 °C night, 60 ± 10% RH). Statistical analysis of the data was made using the ANOVA test and the "a posteriori" comparisons were made using the Tukey-HSD test (both tests with a significance level of 5%). The duration of the development period was longer in treatments where a higher density of females per host was used. When five females per host were used, the mean number of parasitoids that emerged per pupa was higher. The data showed a tendency to a decrease in the amount of parasitoids emerged per host, especially of female, when used high quantities of female per host. Higher parasitism rates were observed in the 3:1 and 5:1 treatments and an increase in the percentage of unviable pupae was observed, probably due to an increase of female densities in the treatments, possibly a consequence of superparasitism.

Identification of infectious agents in onychomycoses by PCR-terminal restriction fragment length polymorphism.

A fast and reliable assay for the identification of dermatophyte fungi and nondermatophyte fungi (NDF) in onychomycosis is essential, since NDF are especially difficult to cure using standard treatment. Diagnosis is usually based on both direct microscopic examination of nail scrapings and macroscopic and microscopic identification of the infectious fungus in culture assays. In the last decade, PCR assays have been developed for the direct detection of fungi in nail samples. In this study, we describe a PCR-terminal restriction fragment length polymorphism (TRFLP) assay to directly and routinely identify the infecting fungi in nails. Fungal DNA was easily extracted using a commercial kit after dissolving nail fragments in an Na(2)S solution. Trichophyton spp., as well as 12 NDF, could be unambiguously identified by the specific restriction fragment size of 5'-end-labeled amplified 28S DNA. This assay enables the distinction of different fungal infectious agents and their identification in mixed infections. Infectious agents could be identified in 74% (162/219) of cases in which the culture results were negative. The PCR-TRFLP assay described here is simple and reliable. Furthermore, it has the possibility to be automated and thus routinely applied to the rapid diagnosis of a large number of clinical specimens in dermatology laboratories.

Revision of the genus Harpasus Mulsant (Coleoptera, Coccinellidae, Chilocorini)

Harpasus Mulsant, 1850 was studied based on the morphology of the exoskeleton and genitalia. The type material of Harpasus aureus Almeida & Carvalho, 2006, H. quadrifolium González, Corrêa & Almeida, 2008 and a homotype of H. zonatus (Mulsant, 1850) were examined. The lectotype of H. eversmanni (Mulsant, 1850) was designated and two new species were described, Harpasus unifasciatus sp. nov. (Teresópolis, RJ, Brazil) and Harpasus ferrugineus sp. nov. (Puerto Carreño, Vichada, Colombia). Herein a diagnosis for the genus and its seven species, identification key and information about biological aspects are presented.

Identification of C(4)-dicarboxylate transport systems in Pseudomonas aeruginosa PAO1.

Pseudomonas aeruginosa utilizes preferentially C(4)-dicarboxylates such as malate, fumarate, and succinate as carbon and energy sources. We have identified and characterized two C(4)-dicarboxylate transport (Dct) systems in P. aeruginosa PAO1. Inactivation of the dctA(PA1183) gene caused a growth defect of the strain in minimal media supplemented with succinate, fumarate or malate, indicating that DctA has a major role in Dct. However, residual growth of the dctA mutant in these media suggested the presence of additional C(4)-dicarboxylate transporter(s). Tn5 insertion mutagenesis of the ΔdctA mutant led to the identification of a second Dct system, i.e., the DctPQM transporter belonging to the tripartite ATP-independent periplasmic (TRAP) family of carriers. The ΔdctA ΔdctPQM double mutant showed no growth on malate and fumarate and residual growth on succinate, suggesting that DctA and DctPQM are the only malate and fumarate transporters, whereas additional transporters for succinate are present. Using lacZ reporter fusions, we showed that the expression of the dctA gene and the dctPQM operon was enhanced in early exponential growth phase and induced by C(4)-dicarboxylates. Competition experiments demonstrated that the DctPQM carrier was more efficient than the DctA carrier for the utilization of succinate at micromolar concentrations, whereas DctA was the major transporter at millimolar concentrations. To conclude, this is the first time that the high- and low-affinity uptake systems for succinate DctA and DctPQM have been reported to function coordinately to transport C(4)-dicarboxylates and that the alternative sigma factor RpoN and a DctB/DctD two-component system regulates simultaneously the dctA gene and the dctPQM operon.

Key to the Australasian and Oceanian genera of Muscidae (Diptera)

A key to 51 Australasian and Oceanian genera of Muscidae is given. The Catalogue of Australasian and Oceanian Diptera and its on line version last reviewed in May 2007 were used as taxonomic guides. Some more recently synonyms and taxonomic changes were updated. For each genus, a brief diagnosis, number of valid species in these regions and comments, when pertinent, are also included. Some morphological diagnostic characteres are illustrated in order to help the use of the key.

Identification of an agrin mutation that causes congenital myasthenia and affects synapse function.

We report the case of a congenital myasthenic syndrome due to a mutation in AGRN, the gene encoding agrin, an extracellular matrix molecule released by the nerve and critical for formation of the neuromuscular junction. Gene analysis identified a homozygous missense mutation, c.5125G>C, leading to the p.Gly1709Arg variant. The muscle-biopsy specimen showed a major disorganization of the neuromuscular junction, including changes in the nerve-terminal cytoskeleton and fragmentation of the synaptic gutters. Experiments performed in nonmuscle cells or in cultured C2C12 myotubes and using recombinant mini-agrin for the mutated and the wild-type forms showed that the mutated form did not impair the activation of MuSK or change the total number of induced acetylcholine receptor aggregates. A solid-phase assay using the dystrophin glycoprotein complex showed that the mutation did not affect the binding of agrin to alpha-dystroglycan. Injection of wild-type or mutated agrin into rat soleus muscle induced the formation of nonsynaptic acetylcholine receptor clusters, but the mutant protein specifically destabilized the endogenous neuromuscular junctions. Importantly, the changes observed in rat muscle injected with mutant agrin recapitulated the pre- and post-synaptic modifications observed in the patient. These results indicate that the mutation does not interfere with the ability of agrin to induce postsynaptic structures but that it dramatically perturbs the maintenance of the neuromuscular junction.

Host preference of the egg parasitoids Telenomus remus and Trichogramma pretiosum in laboratory

Host preference of the egg parasitoids Telenomus remus and Trichogramma pretiosum in laboratory. This research aimed to evaluate the host preference of the egg parasitoids Telenomus remus and Trichogramma pretiosum. Trials were carried out in laboratory, under controlled environmental conditions (25 ± 2ºC temperature; 70 ± 10% RH; and 14 h photophase). The parasitoid searching behavior was evaluated based on the distribution (%) of eggs parasitized by each parasitoid, on egg masses of each host species. Results showed the host preference of T. remus by Spodoptera cosmioides eggs. T. pretiosum, reared in A. gemmatalis eggs, choose to parasitize always eggs of the host where the parasitoid had been reared. The egg preference was not observed when T. pretiosum was reared in S. frugiperda eggs. These results show that, in general, host preference of T. remus is less influenced by the host where it is developed than T. pretiosum. Host preference is an important parameter for biological control programs because more than one pest species may occur in the field, different from those where they were reared in the laboratory.

Identification and Characterization of Natural Anti-Breast Cancer Compound: Costunolide

Breast cancer is the most common cancer among women, 23% (1.3 million) of the total of new cases and the second leading cause of cancer death in women exceeded only by lung cancer. Natural medicines have been proven to be a central source of narrative agents with a pharmaceutical potential. Costunolide is sesquiterpene lactones consisting of diverse plant chemicals that exhibit anti cancer action through cytotoxic effects on various cancer cells. The objectives of present study were to explore the effects of natural compounds on the proliferation of MCF-7 cells and to determine the role of ROS in natural compounds-induced apoptosis in breast cancer cells with a therapeutic potential. Results showed that costunolide screened, possess potent anticancer properties against breast cancer MCF-7 cells, Costunolide was observed as strong anti-proliferative agent with IC50 = 50µM. The anti-proliferative effect of costunolide on MCF cells was confirmed by live/dead assay using fluorescent probes calcein AV/PI. The results demonstrated that treatment of cells with costunolide decreased the viability of MCF-7 cells in a dose-dependent manner. To determine the costunolide-induced apoptosis, flow cytometric analysis was carried out. The results showed that costunolide induced apoptosis in a dose-dependent manner in breast cancer MCF-7cells. ROS are well known mediators of intracellular signaling of cascades. The excessive generation of ROS can induce oxidative stress, loss of cell functioning, and apoptosis. In the present study, we assumed that costunolide might arouse ROS level, which could be involved in induction of apoptosis. Therefore, the intracellular ROS level was measured using the ROS-detecting fluorescence dye 2, 7-dichlorofluorescein diacetate (DCF-DA). Interestingly these effects were significantly abrogated when the cells were pretreated with N-acetyl- cysteine (NAC), a specific ROS inhibitor. Costunolide induces apoptosis through extrinsic pathway in MCF-7 breast cancer cells, In order to examine whether costunolide suppresses cell growth inducing apoptotic cell death, we analyzed DNA contents and apoptosis-related proteins expression level by flow cytometry and western blot, respectively in MCF-7 breast cancer cells we investigated whether costunolide activates extrinsic apoptotic pathway. We examined the expression levels of death receptor signaling-related proteins, caspase-3, and PARP. The results showed that procaspase-3 was cleaved to yield 17 and 20kDa fragments and activation of PARP in treated cells with 25 and 50μM of costunolide. Costunolide induce apoptosis through intrinsic mitochondria pathway in MCF-7 breast cancer Cells. We examined the expression levels of mitochondrial apoptotic pathway related proteins such as anti-apoptotic protein, B-cell lymphoma protein-2 (Bcl2), and pro-apoptotic protein Bax. Costunolide involved in the down regulation of Bcl-2 and up regulation of Bax. These results suggest that costunolide may have beneficial effects for the reduction of breast cancer growth, and new therapeutic strategy for the treatment of human cancers.

Parasitizing behavior of Cervellus piranga Penteado-Dias (Hymenoptera, Braconidae, Braconinae) on papaya borer weevil Pseudopiazurus obesus Marshall (Coleoptera, Curculionidae)

Parasitizing behavior of Cervellus piranga Penteado-Dias (Hymenoptera, Braconidae, Braconinae) on papaya borer weevil Pseudopiazurus obesus Marshall (Coleoptera, Curculionidae). The papaya borer weevil Pseudopiazurus obesus is a pest associated with papaya crops in Brazil and Cervellus piranga is a naturally-occurring parasitoid which may contribute to regulate populations of this pest. We aimed at describing the parasitizing behavior of the parasitoid C. piranga on papaya borer weevil P. obesus larvae under field conditions. The sequence of events related to the parasitizing behavior of C. piranga is similar to other braconid species and includes the location and recognition of the attacked host plant followed by walks on the host plant and touching it with the tip of the antennae and the ovipositor. In the following event, the parasitoid assesses the suitability of the host by speeding up antennae and ovipositor movements. After locating and accepting the host, the parasitoid remains inactive on oviposition aperture sites and starts moving the antennae. Afterwards, the parasitoid inserts its ovipositor and starts cleaning it repeatedly. The female of C. piranga inserts the ovipositor through the hole and lay one egg into the papaya weevil borer. After oviposition, the female cleans continuously both ovipositor and antennae before leaving to forage for a new host.

First report and morphological redescription of Teleonemia morio (Stål) (Hemiptera, Tingidae) in Annona squamosa L. (Annonaceae) in Brazil

First report and morphological redescription of Teleonemia morio (Stål) (Hemiptera, Tingidae) in Annona squamosa L. (Annonaceae) in Brazil. This is the first report of a severe attack of Teleonemia morio (Stål, 1855) (Hemiptera, Heteroptera, Tingidae) on Annona squamosa L. (custard apple), causing up to 80% of losses of infested trees. In order to facilitate the identification of this insect pest, the adult female of T. morio is redescribed based on specimens collected in Palmeira dos Índios, Alagoas, Brazil.

Revision of Central American species of Euryomma Stein (Diptera, Fanniidae), with description of two new species and updates of distributional records

Euryomma is the second largest genus within Fanniidae, but only two species, E. panamensis Chillcott, 1958 and E. rettenmeyeri Chillcott, 1958 have been recorded in Central America. In this paper we describe two new species of the genus found in Costa Rica, Euryomma chillcotti sp. nov. (Province of Puntarenas) and Euryomma steini sp. nov. (Provinces of Cartago and Limón). Euryomma tahami Grisales, Wolff & Carvalho, 2012 is recorded for the first time to Costa Rica. Identification key to all species found in Central America, descriptions and re-descriptions of species occurring in the region and illustrations of the terminalia of new species are presented.